The objective of this work was to determine the behaviour of chalcopyrite (Cp) and pyrrhotite (Po) in microflotation tests using pure minerals and mixtures in the presence of Leptospirillum ferrooxidans. Tests were carried out in the presence and absence of an unconventional collector, isopropyl ethyl thionocarbamate. The results indicate that chalcopyrite and pyrrhotite act differently alone than in a mixture which means the presence of pyrrhotite inhibits chalcopyrite recovery. For chalcopyrite, the flotation rate significantly increases in the presence of bacteria in the first 10 min of contact which is an indication of a first order kinetic equation. The results in the presence of an unconventional collector suggest that the interaction between mineral and thionocarbamate is chemical and not due to the galvanic effect.